小鼠皮肤切创愈合过程中caspase-8表达的研究

目的检测皮肤切创愈合过程中caspase-8的表达,探讨用caspase-8表达的规律推测皮肤损伤时间。方法建立小鼠皮肤切创模型,应用免疫组织化学技术和Western blot方法检测切创后不同时间皮肤中caspase-8的表达,以未切创小鼠皮肤组织作对照。结果伤后12~24h的损伤皮肤组织中部分单核细胞表达caspase-8。随伤后时间延长,caspase-8阳性细胞以单核细胞及成纤维细胞为主。结论伤后3~6h,caspase-8阳性细胞比率较低,12h后逐渐升高,3d达到高峰,以后逐渐下降。实验组中,1、3、5、7d组可见明显的caspase-8活化片段,位于分子量Marker44/42处。结论小鼠皮肤切创愈合过程中,caspase-8在损伤区单核细胞及成纤维细胞中表达,其时序性变化规律可望用于皮肤损伤时间的推断。     

Postmortem wound dehiscence. A report of three cases

Three cases of healing incised wounds that dehisced following death are reported. All three deaths were apparent drownings during summer months. The incised wounds had occurred from 1 week to 6 months prior to death. The length of post-mortem immersion ranged from 2 days to 3 weeks, and all three decedents exhibited significant decompositional changes. Proper recognition of this postmortem artifact eliminates undue suspicions and contributes to proper identification.     

RT-PCR检测不同时间大鼠皮肤切创TGF-β_1mRNA表达

目的 探讨TGF-β_1mRNA在不同时间的表达变化及其与伤后经过时间的关系。方法 应用逆转录-聚合酶链反应(RT-PCR)分别对大鼠生前0.5h、1h、3h、6h、12h、48h、72h、96h、168h和死后0.5h、1h、3h的皮肤切创组织中的TGF-β_1mRNA表达变化进行检测,采用ID软件对凝胶扫描的数据进行密度分析。结果 TGF-β_1mRNA在生前皮肤切创后0.5h上升,3h开始急剧升高,48h含量达到峰值;死后皮肤切创未见TGF-β_1mRNA的表达。结论 大鼠皮肤切创TGF-β_1mRNA的表达具有时间相关性,RT-PCR是检测细胞因子在基因水平表达的灵敏方法。     

Aging of wound healing in an experimental model in mice

Healing of incised wounds in the backs of mice was studied to establish criteria that would allow the timing of stages during cutaneous wound healing. Starting from day 4 after the incisions were made, three mice were killed every 3 days until day 40. Sections from wounds were stained with hematoxylin-eosin, a reticulin procedure, and picrosirius red. Unstained sections were used for blue autofluorescence. The significance of the findings from the ultraviolet study was limited, but the picrosirius red-polarized light procedure was found to be useful for estimating the age of the repair process as polarization colors of greater wavelengths progressively increased with the age of the lesion.     

皮肤切创愈合中caspase-3表达的免疫组化研究

目的探讨皮肤切创损伤愈合过程中,caspase-3在损伤区内的表达以及不同损伤时间caspase-3的变化规律。方法应用免疫组织化学技术对33例不同损伤时间小鼠皮肤切创组织中caspase-3的表达进行研究。同时以3例非切创小鼠皮肤组织做对照。结果伤后6h的损伤皮肤组织中可见少量中性粒细胞表达caspase-3,伤后12～24h,大部分浸润的中性粒细胞及部分单核细胞为caspase-3阳性。随伤后时间延长,caspase-3阳性细胞以单核细胞及成纤维细胞为主。伤后0～3h,caspase-3阳性细胞比率较低,为(4.53±6.53)%,12h后逐渐增加,伤后3d达高峰,为(62.66±4.84)%,其后逐渐下降。结论小鼠皮肤损伤愈合过程中,caspase-3可能在诱导损伤区内中性粒细胞、单核巨噬细胞及成纤维细胞发生凋亡过程中发挥重要作用,同时,caspase-3的规律性表达可用于损伤时间的推断。     

Characteristics of Bone Injuries Resulting from Knife Wounds Incised with Different Forces

The aim of this research was to experimentally determine the characteristics of incised bone wounds, which are commonly found in defense injuries. A specially constructed pivoting arm device was used to inflict wounds with controlled forces and direction. Five knives were selected to inflict the wounds on porcine forelimbs. Eight incised wounds were made per knife per force. A larger knife and a greater force caused longer and wider bone wounds. Comparisons of individual knives at the two forces produced varying results in the bone wounds. A correlation was seen between the force and the length (r = 0.69), width (r = 0.63), and depth (r = 0.57) of bone wounds. Serrated‐edge and nonserrated knives can be distinguished from the appearance of the wound. The outcomes may be applicable in forensic investigations to ascertain the forces associated with incised wounds and identify the specific knife used.     

小鼠皮肤切创愈合过程中caspase-6、-7表达的免疫组织化学研究

目的　观察皮肤切创损伤愈合过程中 ,caspase 6、 7在损伤周边区内的表达及其变化规律。方法　应用免疫组织化学技术观察伤后不同时间小鼠皮肤切创组织中caspase 6、 7的表达 ,以非切创小鼠皮肤组织作对照。 结果　伤后 6h的损伤皮肤组织中可见少量多核粒细胞表达caspase 6、 7,伤后 12～ 2 4h ,大部分浸润的多核粒细胞及部分单核细胞为caspase 6、 7阳性。随伤后时间延长 ,caspase 6、 7阳性细胞以单核细胞及成纤维细胞为主。伤后 0～ 3h ,caspase 6、 7阳性细胞比率较低 ,12h后逐渐增加 ,伤后 3d达高峰 ,其后逐渐下降。 结论　小鼠皮肤损伤愈合过程中 ,caspase 6、 7在损伤周边区内中性粒细胞 ,单核细胞及成纤维细胞中表达 ,其时序性变化规律可望用于皮肤损伤时间的推断。     

A study on the vital reaction in wounded skin: simultaneous determination of histamine and polyamines in injured rat skin by high-performance liquid chromatography

This paper describes a modified method of simultaneous quantitative determination of histamine and polyamines (putrescine, cadaverine, spermidine and spermine) in incised skin wounds of rats using high-performance liquid chromatography (HPLC) with a strong-cation exchange column, shim-pack ISC-05/S0504P, employing a linear gradient of 0.7-2.5 M sodium chloride. There was scarcely any change in the histamine content of the vital wounds throughout the one week experimental period, although content decreased slightly in 1-h-old wounds. On the other hand, concentrations of putrescine and spermidine increased suddenly 12 h after wounding, and spermine content increased slightly in 4-day-old wounds. There was no post-mortem change in the histamine, spermidine and spermine contents of vital wounds allowed to stand at 25 degrees C up to 24 h after death. Based on these results, this HPLC method may be useful for the determination of wound age during the period of healing.     

小鼠皮肤切创愈合过程中iNOS和eNOS表达的免疫组织化学研究

目的观察小鼠皮肤切创愈合过程中,iNOS和eNOS在损伤区及损伤周边区内的表达及其变化规律。方法小鼠背部制作全层切创,应用免疫组织化学技术观察伤后切创组织中iNOS和eNOS的表达,并和无切创的小鼠作为对照。结果损伤区及损伤周边区细胞,伤后3h的损伤皮肤组织中可见少量的多型核细胞表达iNOS和eNOS,伤后6～24h,大部分浸润的中性粒细胞和单核细胞为iNOS和eNOS阳性。随着时间的延长,iNOS和eNOS阳性细胞以单核细胞和成纤维细胞为主。伤后3hiNOS的阳性细胞比率较低,6h～1d持续性增加并于1d达到最高峰,3～7d维持在一个相对稳定的水平直至伤后10d再次达高峰,10～14d开始下降。eNOS的阳性细胞率在伤后1～3h表达较低,6h～3d持续性增高,并在3d达到最高峰,在其后的5d内保持稳定表达,之后开始下降。而且损伤区及损伤周边区、特别是肉芽组织内的新生血管可见iNOS和eNOS不同强度的表达。结论小鼠皮肤切创愈合过程中,iNOS和eNOS在损伤周边区内多型核细胞、单核细胞和成纤维细胞中表达,其时序性变化可望用于皮肤损伤时间的推断。     

Diagnostic ability of D-dimer in the establishment of the vitality of wounds

The potential of skin measurements of D-dimer (DD), a specific derivative of crosslinked fibrin, for the diagnosis of the vital origin of wounds was investigated in 67 human vital skin wounds and homolateral control pieces of skin from 53 autopsies performed at the Institute of Legal Medicine of Coimbra (Portugal) during 1992. Our results demonstrate that DD levels for the incised vital skin wounds (n = 50) were much higher on average than their controls (P < 0.01) which were actually incised injuries produced postmortem at the autopsy. We did not find any statistical difference in average DD levels among the remaining injuries (seven abrasions and ten contusions) in comparison to their respective controls. DD may be used, with certain limitations, as a marker of the vitality of incised wounds. The authors believe this is the first time this marker has been applied in forensic pathology. It is an example of an easy and quick application technique, with the added advantage of low cost.     

Arterial Perfusion—A Useful Technique for Evaluating Incised Wounds

A 54‐year‐old woman was found deceased with incised wounds of both sides of her neck and both wrists. Postmortem CT scanning revealed air in the heart and in the dural veins in continuity with air in the right jugular vein. Death was due to incised wounds of the wrist and neck with blood loss and air embolism. The manner of death was suicide. At autopsy, perfusion of the thoracic ascending aorta produced a fine stream of water emanating from an incised wound of the right ulnar artery with no significant leakage of water from the wound of the left wrist. There was also leakage from the facial artery branch of the right external carotid artery. Perfusion testing can be used as a screening test prior to formal dissection and also to identify small vessels that may not be obvious on standard examination of an exsanguinated field.     

小鼠皮肤切创IL-10和IL-4的表达及其与损伤时间关系

目的　探讨IL 10、IL 4在小鼠皮肤切创愈合过程中的表达变化规律及其与损伤时间的关系。方法　应用免疫组织化学及图像分析技术 ,观察实验小鼠不同时程的生前伤 ( 0 5～ 168h)及死后伤 ( 1～ 6h)皮肤创伤局部IL 10、IL 4的表达情况。结果　生前伤 ,IL 10在伤后 1～ 3h阳性反应逐渐增强 ,3h达峰值 ,2 4h降至较低水平 ,伤后 48h再次升高 ,72h又达新峰值 ,其阳性表达部位主要在表皮细胞及单核 /巨噬细胞 ;IL 4于伤后 2 4h出现明显阳性反应 ,96h表达水平达峰值 ,168h仍维持在较高水平 ,其阳性细胞主要为创伤局部的成纤维细胞。死后伤 ,IL 10仅于死后 1～ 3h呈阳性染色 ,IL 4未见阳性染色。结论　IL 10、IL 4在小鼠皮肤切创愈合过程中的表达呈现一定的时序性变化。     

Expression of adhesion molecules in skin wounds: diagnostic value in legal medicine

The adhesion molecules identified in recent years can help improve the diagnosis of the wound age, especially of injuries with a short survival time. This is also indicative of the vitality of the wounds. The material investigated in the study originated from 465 skin wounds. The samples were taken from human autopsy material, during the surgical treatment of wounds (excision) of patients and from experimental incised wounds of mice. To judge the age of skin wounds the endothelial adhesion molecules were detected in paraffin sections after autoclaving and using the ABC technique. Human skin wounds: strong positive staining was observed of ICAM-1 1.5 h at the earliest and 3.5 days at the latest, for the P-selectin 3 min at the earliest and 7 h at the latest, for the E-selectin 1 h at the earliest and 17 days at the latest and for VCAM-1 3 h at the earliest and 3.5 days at the latest after the time of injury. The L-selectin was expressed constitutively. Mice skin wounds: strong positive immunohistochemical reactions were found as a rule earlier than in human skin wounds. The detection of an increased expression of ICAM-1, VCAM-1 and P- and E-selectins can improve the wound age assessment in injuries with short survival times.     

Homicide by unusual‐edged weapons: Forensic considerations of two cases

Homicide by stab wound is common worldwide. However, the use of unusual edged weapons is rarely reported in the forensic literature. Here, we report two cases of homicide involving a Japanese sword, or katana, and a sickle. Both of these weapons can create either stab or incised wounds with some particularities. In our cases, characteristics of the external wounds, internal injuries, and the depth of penetration raised the possibility of an unusual weapon. Meticulous forensic examination of these injuries (morphological characteristics, depth of penetration, trajectories, and cut marks in the bone) may allow the forensic expert to determine the type of weapon used.     

小鼠皮肤切创愈合过程中caspase-9、-3表达的研究

目的研究caspase-9、-3在小鼠皮肤切创愈合过程中的表达及其变化规律。方法在小鼠背部正中制作皮肤全层切创模型,应用免疫组化染色技术观察切创及切创周边区内caspase-9、-3的表达情况,以无切创的小鼠皮肤作为对照。结果对照组中caspase-9、-3表达于表皮层,毛囊及皮脂腺。伤后3h损伤区及损伤周边区中可见少量多核粒细胞表达caspase-9、-3,6~24h,部分浸润的多核粒细胞和单核细胞caspase-9、-3阳性,此后caspase-9、-3阳性细胞逐渐以单核细胞和成纤维细胞为主。伤后caspase-9、-3阳性细胞率逐渐升高并在3d达到高峰,此后逐渐下降,至14d最低。结论caspase-9、-3可能引发正常小鼠皮肤表皮、毛囊和皮脂腺细胞的凋亡并参与正常皮肤细胞的自我更新;在小鼠皮肤切创愈合过程中,caspase-9、-3在多核粒细胞、单核细胞及成纤维细胞中表达,其时序性变化规律可望用于皮肤切创损伤时间的判定。     

应用ELISA测定成人皮肤切创纤维连接蛋白

应用双抗体夹心酶联免疫吸附剂测定法（ELISA），定量研究了12例成人腹部2小时以内手术切创皮肤可检测的Fn。实验批内变异系数＜5％，批间变异系数＜10％，检测Fn浓度范围3．91～1000ng／ml。结果：成人腹部每克皮肤可检测Fn含量为7．5920±1．7364μg（M±SD）；随着损伤时间延长，不同时间段创伤局部皮肤Fn的含量逐渐升高；各不同时间段与损伤即刻皮肤Fn含量的差值和损伤时间之间存在直线相关（r=0．9843）。经方差分析处理，发现创伤局部皮肤Fn含量在损伤经历30分钟后，有显著增加。本此结果可为推断切创形成的时间提供参考数据。     

小鼠皮肤切创愈合过程中caspase-3的表达及其活性

目的探讨皮肤切创愈合过程caspase-3的表达、激活情况及其活性的变化规律。方法建立小鼠皮肤切创模型后,应用比色法、Western blotting技术对小鼠皮肤切创愈合过程中不同时间段caspase-3的表达、激活情况及其活性进行研究。结果伤后不同时间的caspase-3活性倍增值、caspase-3酶原表达呈现时间规律性变化,并可见caspase-3的激活片段。结论caspase-3可能在皮肤损伤愈合过程中细胞的凋亡中发挥作用,caspase-3可以成为用于皮肤损伤时间推断的指标。     

Secondary combined suicide viewed by the wife]

A 47-year-old man was found dead at the bottom of a wall outward the door of the cellar of his house. Preliminary physical examination at the scene of the death revealed a laceration of the scalp, an incised wound in front of the neck and several stab wounds in the left chest penetrating through the clothing. Questioned by the police his wife told that her husband who suffered from severe depression had inflicted the incised and the stab wounds himself, had made an unsuccessful attempt at self-strangulation and had finally jumped down from the wall beside the stairs declining to the cellar of their house. As the circumstances of the scenario were suspicious and as the pattern of injuries arose doubts concerning self-infliction homicide was taken into consideration as well. Autopsy showed additional hesitation marks adjacent to the left end of the incised wound in front of the neck. The stab wounds of the chest were restricted to the region of the heart and displayed only minimal penetration of the chest-wall, findings characteristic of self-infliction. As a consequence the manner of death could be determined as prolonged suicide, and exact reconstruction was possible with the help of the wife's testimony.     

Symmetrical "mirror-image" injuries and the "chessboard" pattern: useful markers of self-mutilation

The evaluation of injuries due to self-mutilation may be complicated by atypical lesions and fabricated histories of the causative events. Four cases are described where one of the most striking findings was the presence of "mirror-image" injuries and/or a "chessboard" pattern of intersecting parallel lines from self-inflicted incised wounds. Case 1: A 32-year-old man was dead in his burnt-out car, with symmetrical, "mirror-image", circular burns from the car cigarette lighter over his forehead, cheeks, ears, upper and lower arms, the dorsal and palmar aspects of the hands, the anterior chest wall including the nipples, the anterior abdomen, thighs, lower legs, and the back of his neck. The deceased had a history of psychotic depression and had died of burns and inhalation of products of combustion. Case 2: A 19-year-old woman who fabricated a history of assault demonstrated multiple, superficial, "mirror-image" injuries of her thighs and forearms. Case 3: A 32-year-old man who exsanguinated from cut wrists showed multiple, superficial, and "mirror-image" incised wounds of the wrists, with a "chessboard" pattern. Case 4: A 26-year-old man died after jumping from a building. He had multiple superficial, "mirror-image" incised wounds of his anterior chest and thighs, with a typical "chessboard" pattern. Given the difficulties that sometimes arise in determining whether injuries have been self-inflicted or not, the finding of symmetrical "mirror-image" injuries, and/or a "chessboard" pattern formed by 2 groups of regular, parallel, superficial incised wounds running at right angles to each other provides compelling evidence of self-mutilation, unless a plausible alternative explanation is available.     

The 'break enter and die' syndrome may involve significant injury to major neck vessels

Death from incised wounds of major vessels may occur during illegal entry of premises. Wounds are often relatively unimpressive having been caused by thin shards of glass. Alcohol and/or drugs have often been taken that have impaired coordination and contributed to a failure to take appropriate action. A 37-year-old man slipped and fell while attempting to enter a house through a window that he had just broken. He was found dead surrounded by a large amount of blood. At autopsy, a single horizontal, deeply incised wound of the anterior neck was present with transection of the internal jugular veins bilaterally and the right common carotid artery. Toxicological studies were negative for alcohol and common drugs. Death was due to exsanguination and air embolism. This case demonstrates that the injuries sustained during such activities may be quite extensive and involve major neck vessels. Alcohol and drug intoxication are not necessarily involved.